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Ultrastructural cytochemistry of complex carbohydrates in osteoblasts, osteoid, and bone matrix

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Summary

Glycosaminoglycans (GAGs) and glycoproteins are essential components for osteogenesis. We have examined rat osteoblasts, osteoid, transitional zone, and fully calcified bone matrix, utilizing Spicer's high-iron diaminethiocarbohydrazide-silver protein (HID-TCH-SP) method for sulfated glycoconjugates and Thiéry's periodate-TCH-SP (PA-TCH-SP) method for vicinal glycol-containing glycoconjugates. HID-TCH-SP stained cytoplasmic granules of osteoblasts. Stain deposits in the extracellular matrix were observed in decreasing amounts in osteoid, the transitional zone, and fully calcified bone matrix. Enzyme digestion with testicular hyaluronidase removed most HID-TCH-SP stain deposits. PA-TCH-SP staining was observed with increasing intensity in rough endoplasmic reticulum, Golgi saccules, and cytoplasmic granules. Collagen fibrils in osteoid were weakly stained with PA-TCH-SP, and their staining appeared even weaker in fully calcified bone matrix. In contrast, collagen fibrils in calcified cartilage stained intensely with the PA-TCH-SP method. Focal circular profiles (0.1–0.5µm in diameter), which lacked collagen fibrils but reacted moderately with PA-TCH-SP, were frequently seen in the transitional zone and fully calcified bone matrix, but were only occasionally present in osteoid. The presence of testicular hyaluronidase-resistant GAG and acid phosphatase in these focal areas suggests that they represent sites of GAG degradation. The eventual loss of HID-TCH-SP staining in the bone matrix suggests that removal of sulfated glycoconjugates may be a requisite for expansion of initial calcification sites and/or complete calcification.

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Authors and Affiliations

  1. From the Institute of Dental Research and the Department of Anatomy, Nihon University School of Dentistry, Tokyo, Japan

    Minoru Takagi

  2. From the Institute of Dental Research and the Department of Pediatrics and Pathology, The University of Alabama in Birmingham, 35294, Birmingham, Alabama, Tokyo, Japan

    Richard T. Parmley

  3. From the Institute of Dental Research and the Department of Anatomy, Nihon University School of Dentistry, Tokyo, Japan

    Yoshihisa Toda

  4. From the Institute of Dental Research, Tokyo, Japan

    Francis R. Denys

Authors
  1. Minoru Takagi
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  2. Richard T. Parmley
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  3. Yoshihisa Toda
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  4. Francis R. Denys
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Takagi, M., Parmley, R.T., Toda, Y. et al. Ultrastructural cytochemistry of complex carbohydrates in osteoblasts, osteoid, and bone matrix. Calcif Tissue Int 35, 309–319 (1983). https://doi.org/10.1007/BF02405052

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